Engineering an Enhanced EGFR Engager: Humanization of Cetuximab for Improved Developability

Abstract

The epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase whose proliferative effects can contribute to the development of many types of solid tumors when overexpressed. For this reason, EGFR inhibitors such as cetuximab can play an important role in treating cancers such as colorectal cancer and head and neck cancer. Cetuximab is a chimeric monoclonal antibody containing mouse variable regions that bind to EGFR and prevent it from signaling. Although cetuximab has been used clinically since 2004 to successfully control solid tumors, advances in protein engineering have created the opportunity to address some of its shortcomings. In particular, the presence of mouse sequences could contribute to immunogenicity in the form of anti-cetuximab antibodies, and an occupied glycosylation site in FR3 can contribute to hypersensitivity reactions and product heterogeneity. Using simple framework graft or sequence-/structure-guided approaches, cetuximab was humanized onto 11 new frameworks. In addition to increasing humanness and removing the VH glycosylation site, dynamic light scattering revealed increases in stability, and bio-layer interferometry confirmed minimal changes in binding affinity, with patterns emerging across the humanization method. This work demonstrates the potential to improve the biophysical and clinical properties of first-generation protein therapeutics and highlights the advantages of computationally guided engineering.

Development of nanobodies as theranostic agents against CMY-2-like class C β-lactamases

“…Actually, one strategy in view to minimize the size of inhibitors consists in the development of small peptides by peptidomimetics. These present several advantages: (I) an easier production in large scale-up with a cheaper cost, (II) the low tissues penetration of large molecules and antibodies rendering less efficient the drug delivery and its action and (III) the humanization of therapeutics antibodies which can be laborious and which can lead finally to the development of human anti-mouse antibody (HAMA) (44, 45).…”

Assessing developability early in the discovery process for novel biologics

“…Among the antibodies chosen for experimental testing, the computational developability assessments can help identify potential lead molecules that possess an optimal combination of functional as well as physicochemical attributes. However, depending on peculiarities of individual drug discovery projects, these lead molecules may need to be affinity matured, 85 , 86 humanized, 87–89 formatted in novel molecular constructs, 90 and further optimized for fitness with development platforms. This calls for the use of computational protein engineering tools to optimize the lead molecules.…”

Recent Advancements, Limitations, and Future Perspectives of the use of Personalized Medicine in Treatment of Colon Cancer

“…It is a chimeric monoclonal IgG1 antibody of the epidermal growth factor receptor (EGFR). 124 It selectively binds to the extracellular region of the EGFR and blocks the domain from binding to epidermal growth factor (EGF) and other ligands. 125 This blockage results in cell growth inhibition, increased apoptosis of tumor cells, reduced angiogenesis, and metastasis.…”